title: High-throughput screening of one-bead-one-compound peptide libraries using intact cells.
abstract: Screening approaches based on one-bead-one-compound (OBOC) combinatorial libraries have facilitated the discovery of novel peptide ligands for cellular targeting in cancer and other diseases. Recognition of cell surface proteins is optimally achieved using live cells, yet screening intact cell populations is time-consuming and inefficient. Here, we evaluate the Complex Object Parametric Analyzer and Sorter (COPAS) large particle biosorter for high-throughput sorting of bead-bound human cell populations. When a library of RGD-containing peptides was screened against human cancer cells that express αvβ3 integrin, it was found that bead-associated cells are rapidly dissociated when sorted through the COPAS instrument. When the bound cells were reversibly cross-linked onto the beads, however, we demonstrated that cell/bead mixtures can be sorted quickly and accurately. This reversible cross-linking approach is compatible with matrix-assisted laser desorption ionization time-of-flight mass spectrometry-based peptide sequence deconvolution. This approach should allow one to rapidly screen an OBOC library and identify novel peptide ligands against cell surface targets in their native conformation.
authors.affiliation: Translational Prostate Cancer Research Group, University of Alberta , 5-142C Katz Group Building, 114th Street and 87th Avenue, Edmonton, AB, T6G 2E1 Canada.
authors.forename: Choi-Fong
authors.initials: CF
authors.lastname: Cho
authors.forename: Babak
authors.initials: B
authors.lastname: Behnam Azad
authors.forename: Leonard G
authors.initials: LG
authors.lastname: Luyt
authors.forename: John D
authors.initials: JD
authors.lastname: Lewis
mesh_terms: D000595:Amino Acid Sequence; D001331:Automation; D011211:Electric Power Supplies; D057166:High-Throughput Screening Assays / Q000379:methods*; D019151:Peptide Library; D010455:Peptides / Q000737:chemistry*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
chemical_list: D019151:Peptide Library; D010455:Peptides